First report for the resynthesis of Brassica napus by recombining A and C genome from B. juncea and B. carinata , respectively. Also documents B genome introgressions in resynthesized B. napus. Resynthesis of Brassica napus (AACC) was achieved by hybridizing Brassica juncea (AABB) with Brassica carinata (BBCC). This was facilitated by spontaneous chromosome doubling in the F1 hybrid (ABBC) to yield octaploid (AABBBBCC), elimination of extra B genome chromosomes in the resulting octaploid and in subsequent selfed generations, aided with directed selection for fertile plants having B. napus morphology. Twenty-five plants with varying degrees of resemblance to natural B. napus were identified from 17 A5 progenies and assayed for cytogenetic stability and genetic diversity. Majority of these plants, except six (2n = 38) were hyperploids (2n = 40-56). The six plants with 2n = 38 were designated as derived B. napus types. These showed an expected meiotic configuration of 19II at metaphase-I, with 19-19 distribution at anaphase-I. Genotyping based on A and C genome specific primers confirmed genetic identity of six derived (2n = 38) B. napus plants with natural types whereas genotyping with B genome specific primers indicated introgression of B genome segments. This was also confirmed by genomic in situ hybridization (GISH). Strong signals of B genome probe were detected, proving hitherto unreported genetic exchanges between B and A/C chromosomes. These introgressions possibly occurred en route five generations of selfing. Derived plants yielded fertile hybrids in crosses with natural B. napus var. GSC 6. The selfed derived plants as evaluated in A6 plant to progeny rows were morphologically similar to natural B. napus, and meiotically stable. Agronomic assessment of these progenies revealed variation for key morpho-physiological traits. Of special interest were the progenies with plants having oil content exceeding 47% as against about 39-41% in existing cultivars.
Brassica napus introgression lines (ILs), having B-genome segments from B. carinata, were assessed genetically for extent of introgression and phenotypically for siliqua shatter resistance. Introgression lines had 7–9% higher DNA content, were meiotically stable, and had almost normal pollen fertility/seed set. Segment introgressions were confirmed by fluorescent genomic in situ hybridization (fl-GISH), SSR analyses, and SNP studies. Genotyping with 48 B-genome specific SSRs detected substitutions from B3, B4, B6, and B7 chromosomes on 39 of the 69 ILs whereas SNP genotyping detected a total of 23 B-segments (≥3 Mb) from B4, B6, and B7 introgressed into 10 of the 19 (C1, C2, C3, C5, C6, C8, C9, A3, A9, A10) chromosomes in 17 ILs. The size of substitutions varied from 3.0 Mb on chromosome A9 (IL59) to 42.44 Mb on chromosome C2 (IL54), ranging from 7 to 83% of the recipient chromosome. Average siliqua strength in ILs was observed to be higher than that of B. napus parents (2.2–6.0 vs. 1.9–4.0 mJ) while siliqua strength in some of the lines was almost equal to that of the donor parent B. carinata (6.0 vs.7.2 mJ). These ILs, with large chunks of substituted B-genome, can prove to be a useful prebreeding resource for germplasm enhancement in B. napus, especially for siliqua shatter resistance.
C genome chromosome substitution lines of B. juncea constitute a key genetic resource for increased genetic diversity and hybrid performance. C genome chromosome substitution lines were found in the progenies of derived B. juncea (2n = 36; AABB), synthesized through hybridization between B. napus and B. carinata. These were originally recognized based on the morphology and genomic in situ hybridization. Genotyping using the Brassica Illumina 60K Infinium SNP array confirmed the presence of C genome chromosomes in a large number of derived B. juncea genotypes. Three whole chromosome substitutions and 13 major C genome fragment substitutions were identified. Fragment substitutions were primarily terminal, but intercalary substitution(s) involving chromosome C1 and C2 were identified in three genotypes. The size of substituted C genome fragments varied from 0.04 Mbp (C1) to 64.85 Mbp (C3). In terms of proportions, these ranged from 0.10 % (C1) to 100 % (C1, C3 and C7) of the substituted chromosome. SSR genotyping with B genome specific primers suggested that substituting C genome chromosome(s) are likely to have replaced B genome chromosome(s). C1 was the most common substituting chromosome while substituted B chromosome seemed random. Study of the phenotypic traits underlined the importance of the substitution lines (especially of chromosome C1) for conferring superior trait performance (main shoot length and pods on the main shoot). High heterosis was also indicated in hybrid combinations of substitution lines with natural B. juncea. These substitution genotypes constituted a valuable resource for targeted gene transfer and QTL identification.
No abstract
One of the breeding goals in Brassica napus has been to facilitate introgression of novel traits from wild or closely related species through inter- and intra-specific crosses. The present study is based on the evaluation of B. napus – carinata introgression lines (ILs) for introgressed morphological and genetic diversity in comparison to the parental B. napus lines. A set of 81 Brassica napus – carinata ILs, developed previously following a back cross strategy, was assessed for introgressed genetic diversity by comparing them with the 15 B. napus recipient parents under two environments (E1: timely sown and E2: late sown) for phenotypic expression of 13 morpho-physiological traits. Trait averages as well as trait variations were generally higher for ILs, indicating introgression of variability in the desired direction. In comparison to the parents, the ILs were observed to be early to flower, early to mature, higher yielding, had more siliquae on the main axis and the siliquae were longer. ANOVA revealed highly significant differences amongst genotypes for 10 traits of which 4 traits, viz., days to 50% flowering, days to 100% flowering, plant height and pod length were highly significant in both the environments. SNP-based chromosome-wise gene diversity, as estimated using software SELECTIONTOOL, indicated high estimates for individual chromosomes in ILs as compared to the parental lines. The phenotypic variability estimated for majority of the traits in ILs is being utilized for developing new high yielding rapeseed varieties.
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